ultraviolet irradiation o'f DT resistance in cultures of the transformed, xeroderma pigmentosum complementation group-A cell line XP20S(SV40), <strong>and</strong> of the Chinese hamster ovary ~CHO) repair-defective mutant 43-3B . In both cultures, there was a do~e-dependent increase in the frequency of toxinresistant cells . On an equal-dose basis, higher frequencies were observed in the repair-deficient cell lines than in their respective repair-proficient controls, GM637(SV40) <strong>and</strong> CHO-9 . At equal levels of survival, however, the frequencies of DTs cells were similar in the repair- defi~ient <strong>and</strong> proficient cells . These result provide further evidence that the DT" cells detected by the autoradiographic assay are indeed mutants . They also indicate a simple method of studying various aspects of mutation in cells that, because of e .g . defective DNA-repair, are hard to clone <strong>and</strong> use in a colony assay . (Work supported by the Israel Cancer Association .) 583 SOFTWARE DEVELOPMENT FOR MICRONUCLEUS (MN) SCORING . R .R . Tice, J .T . MacGre9or*, C . Campfield, A . Pellom, L . Williams** <strong>and</strong> C .H . Nauman**, Integrated Laboratory Systems, PO Box 13501, Res . Tri . Park, NC 27709, *Toxicology Consulting Services . Inc ., 383 Diablo Rd, Suite 100, Danville, CA 94526, <strong>and</strong> **EPA/EMSL, P0 Box 89193, Las Vegas, NV 89193 . Based on input from a panel of experts in the field, a software program, suitable for an IBM compatible PC, has been developed for the collection <strong>and</strong> analysis of micronuclei data obtained from in vtvo test systems . Experimental design information <strong>and</strong> MN data obtained on a number of cell types are easily entered using a series of menus, statistically analyzed by a variety of statistical models <strong>and</strong> presented both in tabular <strong>and</strong> graphic form . In the statistical analysis, the data are evaluated for scorer, sex, treatment, sample time, <strong>and</strong> animal effects . The use of this software will help to st<strong>and</strong>ardize in vivo MN data analysis <strong>and</strong> presentation . Although the research described in this article has been supported by the U .S . EPA through contract number 68-C8-0068 to Integrated Laboratory Systems, it has not been subjected to Agency review <strong>and</strong> therefore does not necessarily reflect the views of the Agency <strong>and</strong> no official endorsement should be inferred . . 584 • ANALYSIS OF MOUSE MICRONUCLEI BY HIGH SPEED FLOW CYTOMETRY . A. M . Tometsko, Litron Laboratories, Rochester, N. Y . (USA) The mouse micronucleus assay is widely used to evaluate the clastogenic activity of chemicals . The essay involves scoring the number of cells containing micronuclei (MNS) In either blood or bone marrow preparations using fluorescent or bright field microscopy . The conventional assay is labor intensive end tedlus, <strong>and</strong> requires many hours of microscopic examination to complete an assay. High speed flow cytometry provides the means for analyzing fluorescent cells as they pass through a laser beam at 2,000 cell per second <strong>and</strong> permits the analysis of 30-50 times more cells then manual screening tn e shorter time. For FCM analysis, the cells are stained with fluorescent dyes which effectively label cellular nucleic acids (e .g. MNs) . Experiments will be presented which highlight the distribution of micronucleated cells in peripheral blood of normal <strong>and</strong> ctasto0en treated mice. The location of MNs in different blood cell populations will be shown <strong>and</strong> will provide en indicetion of the feasibility of developing an automated FCM based analysis protocol• The distribution of MNcells will be presented using histograms <strong>and</strong> bivariate graphs. Correlations will be made between manual scoring <strong>and</strong> analysis by high speed flow cytometry . 585 COMPARISON OF IN VIVO SOMATIC CELL MUTATION, CHROlie/SOME ABERRATION, SISTER CHROMATID EXCHANGE, MICRONUCLEI FORMATION AND URINE MUTAGENICITY IN STEEL FOUNDRY WORKERS . D .J . Tomkins, D .R . McCalla, <strong>and</strong> E .S . Gibson, McMaster University <strong>and</strong> Dofasco Inc .,Hamilton, Ontario, Canada . Preliminary results of genetic toxicologic monitoring of 125 steel foundry workers with a higher rate of lung cancer have been reported (Environ . Mutag . 7(S .3) : 33,1985) . Chromosome aberration (CA), sister chromatid exchange (SCE), <strong>and</strong> morning <strong>and</strong> afternoon urine mutagenicity (MUTAM,MUTPM), but not micronuclei (HN), were all significantly elevated with smoking (p- .001, .004, .008, .005 respectively), but not with occupation . The somatic cell mutation test (SCMT) has now been completed for 37 workers <strong>and</strong> the statistical analysis did not show any significant effects of smoking or occupation . In the case of CA, there was a significant interaction between smoking <strong>and</strong> occupation, particularly when gaps were not included in the total number of aberrations per cell (p- .01) . When specific types of abnormalities were analyzed separately, breaks appeared to be the moat important contributors to the effect of the interaction . Two http://legacy.library.ucsf.edu/tid/clb93d00/pdf 1989 EMS Abstracts 201 Notes
202 1989 EMS Abstracts Notes important conf.unieta -were identified : age was positively correlated with CA, !IN <strong>and</strong> . . ° .-aelayed/proloapediEWl'division in 48 hr culturas, <strong>and</strong> drug exposure significantly affected IiUTPW<strong>and</strong> CA . The different genetic tests were often correlated with each other . In partic . ,HUTAH was positively correlated with CA (p- .02), MUT'PN with CA, SCE, <strong>and</strong> SCH1 , .04, .01 respectively) . On the other h<strong>and</strong>, SCE <strong>and</strong> SCMf may have been negatively correlated (p- .05), <strong>and</strong> there was no correlation between CA, SCE <strong>and</strong> ALN . This suggests that urine mutagenicity may measure the metabolic products of a genotoxic exposure, but that the types of genetic damage detected by the lymphocyte tests may vary from subject to subject . http://legacy.library.ucsf.edu/tid/clb93d00/pdf MAMMALIAN DNA LIGASE I : THE MOLECULAR DEFECT IN BLOOI,fS SYNDROME Alan E . Tomidnson, Dena D. Lasko, Ams E . WNia <strong>and</strong> Tarnas Lirqalt rnperml Caneer Research Fund, Clare Hall Laborabries, South t,6mms, Herts ., EN6 SLD, U.K. Mammalian cells contain two clistinct DNA i0ases. The major aclivity In proUteratkp oeNS <strong>and</strong> therefore prohabfy the enzyme involved in DNA replioation Is the NOh mdealar weipAt enzyme. DNA Npue I. A 1dOkd brm of frs enzyme has been purilkd b ~ ope~ ~~ thymus <strong>and</strong> tunan oeos . Theae en:ymes tww been characwised <strong>and</strong> poytbrol antlbod~e ses talaed apakrot DNA Ipases from the bede:bphapea T4 <strong>and</strong> T7, yeasts.J.wevelas <strong>and</strong><strong>and</strong> vaccM4a vkus tonlain conserved repions of amrw add sequence . Poydonai anbbodes have a t sidue peptlde representing one athese conserved raqons <strong>and</strong> these anobot
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